引物延伸预扩增法应用于β-地中海贫血患者胚胎植入前遗传学诊断——附一例报告  

Successful preimplantation genetic diagnosis for beta-thalassemia using primer extension preamplification

作　　者：焦泽旭[1] 庄广伦[1] 周灿权[1] 舒益民[1] 李洁[1] 梁晓燕[1] 张敏芳[1] 邓明芬[1] 

机构地区：[1]中山大学附属第一医院妇产科生殖医学研究中心,广州510080

出　　处：《中华妇产科杂志》2003年第3期143-146,共4页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的　探讨对 β 地中海贫血进行胚胎植入前遗传学诊断 (PGD)的方法。 方法　两对夫妇双方均分别为密码子 41 42 ( TCTT)及插入序列 (IVS) Ⅱ 65 4(C→T)突变杂合子 ,在本中心进行超排卵和体外受精 胚胎移植治疗 ,胚胎活检后应用全基因组扩增技术及反向点杂交进行胚胎PGD ,根据诊断结果选择健康的胚胎移植入子宫。结果　 2例患者共获卵 3 5个 ,受精率为 87% ,共活检胚胎16个 ,获卵裂球 2 5个 ,单卵裂球总扩增效率为 84% ,等位基因脱扣率为 15 %。 2例患者共移植 5个胚胎 ,1例获得妊娠 ,已分娩健康双胎。结论　应用引物延伸预扩增技术可对 β 地中海贫血进行PGD 。Objective To achieve preimplantation genetic diagnosis(PGD) of the couples at risk of having children with beta thalassemia,as an alternative to prenatal diagnosis Methods Two couples carrying different thalassemia mutations of codon 41/42 and codon intervening sequence 2 position 654 received standard in vitro fertilization treatment and intracytoplasmic sperm injection,embryo biopsy and the whole genome was amplified by primer extension preamplification (PEP).Nested polymerase chain reaction was then used to amplify two mutation sites separately. Both were detected by reverse dot blot Results A total of 35 oocytes were retrieved from the two patients. Among them,87% showed two pronuclei,and embryo biopsy was performed on 16 of these embryos and 25 blastomeres were obtained. The amplification efficacy was 84%.The genotype study of non transferred and surplus embryos showed 15% of allele drop out rate. Five embryos were transferred to the uterus of both patients. One pregnancy achieved,resulted in live healthy twin births,which confirmed the results of PGD Conclusions This unaffected pregnancy resulting from PGD by PEP for beta thalassemia demonstrates that this technique can be a effective diagnostic tool for carrier couples who desire a healthy child.

关 键 词：Β-地中海贫血 患者 PGD 遗传学诊断 全基因组扩增 胚胎植入 治疗 密码子 卵裂球 引物 

分 类 号：R714.5[医药卫生—妇产科学]